Metabolic analysis

1. The engineering of metabolic pathways requires the analysis of transgenic organisms, and a specialised facility has been established at the OpenPlant Lab Norwich.

2. Specialised instruments have been acquired in order to complement existing equipment and provide a comprehensive suite of high resolution analytical instruments for metabolite analysis in microbes and plants.

Digital droplet PCR analysis

Digital droplet PCR (ddPCR) converts the exponential, analogue nature of PCR into a linear, digital signal allowing accurate estimations of relative abundance of specific sequences. A Biorad QX100 ddPCR machine will be acquired for the development and assessment of precision editing tools. It will also improve the speed of research utilising traditional transgenic and genetic approaches by allowing swift and accurate assessment of copy number and zygosity.

Q-Extractive MALDI mass spectrometer

This instrument extends the accurate mass identification of novel products and will enable image metabolites in plant cells, so informing engineering for product optimisation. To date we have been limited to a restricted range of chemicals that can be imaged by mass spectrometry. MALDI imaging is not specific to particular metabolites and so is very versatile, providing the potential to extend this ground-breaking approach to the whole gamut of plant products, natural and manipulated.

GC-GC mass spectrometer with sample handling

This instrument provides a range of new capabilities. Natural products such as terpenes have vast diversity of carbon backbones. Structure determines biological activity, and minor changes to synthetic enzymes can have large knock-on consequences for the structures synthesised. GCxGC can tease apart the complexity. Our existing systems can handle only liquid samples; the new inlet system will be able to handle a wide range of different injection methods (e.g. Thermal desorption, head-space and SPME), allowing better investigation of volatile products (e.g. related to market opportunities in flavours and fragrances).

Advion bench-top mass spectrometer

The Advion compact mass spectrometer will allow rapid analysis of TLC plates and column fractions, and be compatible with online use of our current range of HPLC and flash chromatography systems. This instrument will increase throughput, decrease waiting-time for results, and reduce demand on the main platform instruments, freeing them for more sophisticated analyses. This will provide a step-change for (non)natural product isolation and take pressure off the main platform LC-MS instruments.

Bruker Microflex LT MALDI-TOF mass spectrometer

While LC-MS and GC-MS provide good coverage between them, some important areas of natural product chemistry remain difficult in both techniques. One area is glycosylation. Only smaller oligosaccharides can be reliably separated and analysed with existing LC-MS and GC-MS equipment, and a MALDI system will extend coverage to basic glycomics and glycoproteomics applications. Although some of this work – the higher end analyses - could be carried out on the Q-Exactive/MALDI system, the Orbitrap approach has limited m/z range, and will be heavily committed to imaging work.The Microflex LT is an entry-level, bench-top high-performance time-of-flight mass spectrometer that is particularly suited to routine or walk-up quality control and screening applications.